[1]王中超,何延忠,赵栋梁,等.丹参通络解毒汤对博莱霉素诱导的肺纤维化小鼠肿瘤坏死因子样弱凋亡诱导因子/成纤维细胞生长因子诱导14通路的影响[J].新乡医学院学报,2021,38(12):1101-1112.[doi:10.7683/xxyxyxb.2021.12.001]
　WANG Zhongchao,HE Yanzhong,ZHAO Dongliang,et al.Effect of Danshen Tongluo Jiedu decoction on tumor necrosis factor-like weak inducer of apoptosis/fibroblast growth factor-inducible 14 pathway in mice with pulmonary fibrosis induced by bleomycin[J].Journal of Xinxiang Medical University,2021,38(12):1101-1112.[doi:10.7683/xxyxyxb.2021.12.001]
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丹参通络解毒汤对博莱霉素诱导的肺纤维化小鼠肿瘤坏死因子样弱凋亡诱导因子/成纤维细胞生长因子诱导14通路的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
38
期数:
2021年12
页码:
1101-1112
栏目:
基础研究
出版日期:
2021-12-05

文章信息/Info

Title:
Effect of Danshen Tongluo Jiedu decoction on tumor necrosis factor-like weak inducer of apoptosis/fibroblast growth factor-inducible 14 pathway in mice with pulmonary fibrosis induced by bleomycin
作者:
王中超; 何延忠; 赵栋梁; 周　淼
（河南中医药大学第三附属医院肺病科，河南　郑州　450003）
Author(s):
WANG Zhongchao; HE Yanzhong; ZHAO Dongliang; ZHOU Miao
(Department of Pulmonary Diseases,the Third Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450003,Henan Province,China)
关键词:
肺纤维化; 丹参通络解毒汤; 博莱霉素; 肿瘤坏死因子样弱凋亡诱导因子; 成纤维细胞生长因子诱导14
Keywords:
pulmonary fibrosis; Danshen Tongluo Jiedu decoction; bleomycin; tumor necrosis factor-like weak inducer of apoptosis; fibroblast growth factor-inducible 14
分类号:
R285.5；R563
DOI:
10.7683/xxyxyxb.2021.12.001
文献标志码:
A
摘要:
目的　观察丹参通络解毒汤对博莱霉素诱导的肺纤维化小鼠肿瘤坏死因子样弱凋亡诱导因子（TWEAK）/成纤维细胞生长因子诱导14（Fn14）通路的影响，探讨其对小鼠肺纤维化的影响及机制。方法　取C57B/L小鼠50只随机分为空白组、模型组、1，25-二羟维生素D3［1，25（OH）2D3］组、丹参通络解毒汤组、丹参通络解毒汤+TWEAK/Fn14抑制剂 PDL192组。模型组、1，25（OH）2D3组、丹参通络解毒汤组、丹参通络解毒汤+PDL192组小鼠气管内注射博莱霉素建立小鼠肺纤维化模型。造模成功后，空白组和模型组小鼠每日灌胃生理盐水10 mL·kg-1；1，25（OH）2D3组小鼠每日经腹腔注射1，25（OH）2D3 5 μg·kg-1；丹参通络解毒汤组小鼠每日灌胃丹参通络解毒汤汤剂0.039 mL·g-1（按成人60 kg体质量计算，相当于生药15.65 g·kg-1）；丹参通络解毒汤+PDL192组小鼠每日灌胃丹参通络解毒汤汤剂0.039 mL·g-1，并经腹腔注射PDL192 3 mg·kg-1；各组小鼠均每日给药1次，连续给药28 d；末次给药后12 h测体质量，然后经腹腔注射戊巴比妥钠进行麻醉、固定，采集肺支气管肺泡灌洗液（BALF），开胸取肺组织，测肺质量，计算小鼠肺系数。采用酶标仪检测5组小鼠BALF中白细胞介素1β（IL-1β）、肿瘤坏死因子-α（TNF-α）、白细胞介素4（IL-4）水平。苏木精-伊红染色和Masson三色法染色法观察5组小鼠肺组织病理学变化。实时荧光定量聚合酶链式反应法检测5组小鼠肺组织TWEAK、Fn14、转化生长因子β1（TGF-β1） mRNA表达。Western blot法检测5组小鼠肺组织炎症细胞因子TNF-α、IL-1β、细胞间黏附分子（ICAM-1）、血管细胞黏附分子1（VCAM-1）及肺组织TWEAK/Fn14通路因子TWEAK、Fn14、TGF-β1、α-平滑肌肌动蛋白（α-SMA）、母亲信号蛋白同源物3（Smad3）、磷酸化母亲信号蛋白同源物3（p-Smad3）、信号传导和转录激活因子3（STAT3）、酪氨酸磷酸化STAT3（p-STAT3）蛋白相对表达量。结果　空白组小鼠肺组织结构正常，无结缔组织增生，无明显胶原沉积；模型组小鼠肺组织基本形态结构被破坏，支气管结构较为模糊，部分支气管上皮细胞坏死，间质周围炎症细胞浸润，肺组织有大量的胶原沉积；1,25（OH）2D3组小鼠肺组织基本形态结构被破坏，上皮细胞坏死，坏死区域肺泡腔较为狭窄，内含大量细胞碎片和炎症细胞，部分区域可见成纤维细胞增生，胶原沉积明显减少；丹参通络解毒汤组小鼠部分区域肺组织结构被破坏，支气管结构较为完整清晰，部分肺泡结构模糊，肺泡腔狭窄，间质及周围未见水肿或炎症细胞浸润，胶原沉积明显减少；丹参通络解毒汤+PDL192组小鼠肺组织结构较为正常，各级支气管结构完整清晰，肺泡结构较为清晰，肺泡腔内可见少量炎症细胞散在分布，间质及周围未见水肿或炎症细胞浸润，胶原沉积明显减少。空白组、模型组、1,25（OH）2D3组、丹参通络解毒汤组、丹参通络解毒汤+PDL192组小鼠体质量比较差异均无统计学意义（P>0.05）。与空白组比较，模型组、1,25（OH）2D3组、丹参通络解毒汤组小鼠肺系数显著增高（P<0.05）；与模型组比较，丹参通络解毒汤组、丹参通络解毒汤+PDL192组小鼠肺系数显著降低（P<0.05）；与1,25（OH）2D3组比较，丹参通络解毒汤+PDL192组小鼠肺系数显著降低（P<0.05）；1,25（OH）2D3组、丹参通络解毒汤+PDL192组小鼠肺系数与丹参通络解毒汤组比较差异无统计学意义（P>0.05）。与空白组比较，模型组、1,25（OH）2D3组、丹参通络解毒汤组小鼠BALF中IL-1β、TNF-α、IL-4水平显著升高（P<0.05），丹参通络解毒汤+ PDL192组小鼠BALF中IL-1β水平显著升高（P<0.05）；与模型组比较，1,25（OH）2D3组、丹参通络解毒汤组、丹参通络解毒汤+PDL192组小鼠BALF中IL-1β、TNF-α、IL-4水平显著降低（P<0.05）；与1,25（OH）2D3组比较，丹参通络解毒汤组、丹参通络解毒汤+PDL192组小鼠BALF中IL-1β、IL-4水平显著降低（P<0.05）；与丹参通络解毒汤组比较，丹参通络解毒汤+ PDL192组小鼠BALF中IL-1β、IL-4水平显著降低（P<0.05）。与空白组比较，模型组小鼠肺组织TWEAK、Fn14、TGF-β1 mRNA相对表达量及1,25（OH）2D3组小鼠肺组织TWEAK mRNA相对表达量显著升高（P<0.05）；与模型组比较，丹参通络解毒汤组小鼠肺组织Fn14 mRNA相对表达量及丹参通络解毒汤+PDL192组小鼠肺组织TWEAK、Fn14、TGF-β1 mRNA相对表达量显著降低（P<0.05）。1,25（OH）2D3组、丹参通络解毒汤组、丹参通络解毒汤+PDL192组小鼠肺组织TWEAK、Fn14、TGF-β1 mRNA相对表达量各组间比较差异无统计学意义（P>0.05）。与空白组比较，模型组小鼠肺组织TNF-α、IL-1β、ICAM-1、VCAM-1蛋白相对表达量及1,25（OH）2D3组、丹参通络解毒汤组小鼠肺组织TNF-α、IL-1β蛋白相对表达量显著升高（P<0.05）；与模型组比较，丹参通络解毒汤组、丹参通络解毒汤+PDL192组小鼠肺组织TNF-α、IL-1β蛋白相对表达量显著降低（P<0.05）；与1,25（OH）2D3组比较，丹参通络解毒汤组和丹参通络解毒汤+PDL192组小鼠肺组织IL-1β蛋白相对表达量显著降低（P<0.05）；与丹参通络解毒汤组比较，丹参通络解毒汤+ PDL192组小鼠肺组织IL-1β蛋白相对表达量显著降低（P<0.05）。与空白组比较，模型组小鼠肺组织中TWEAK、Fn14、TGF-β1、α-SMA、p-Smad3、STAT3、p-STAT3蛋白相对表达量显著升高（P<0.05），1,25（OH）2D3组小鼠肺组织中TWEAK、TGF-β1、α-SMA、p-Smad3、STAT3、p-STAT3蛋白相对表达量显著升高（P<0.05），丹参通络解毒汤组、丹参通络解毒汤+ PDL192组小鼠肺组织中TWEAK、α-SMA、p-Smad3、STAT3蛋白相对表达量显著升高（P<0.05）；与模型组比较，1,25（OH）2D3组小鼠肺组织p-Smad3蛋白相对表达量显著降低（P<0.05），丹参通络解毒汤组、丹参通络解毒汤+PDL192组小鼠肺组织TWEAK、Fn14、TGF-β1、α-SMA、p-Smad3、STAT3蛋白相对表达量显著降低（P<0.05）。丹参通络解毒汤组、丹参通络解毒汤+PDL192组小鼠肺组织Fn14、TGF-β1、α-SMA、Smad3、p-Smad3、p-STAT3蛋白相对表达量及丹参通络解毒汤小鼠肺组织TWEAK、STAT3蛋白相对表达量与1,25（OH）2D3组比较差异无统计学意义（P>0.05）；丹参通络解毒汤+ PDL192组小鼠肺组织中TWEAK、STAT3蛋白相对表达量与1,25（OH）2D3组比较显著降低（P<0.05）。结论　丹参通络解毒汤能够减轻博莱霉素诱导的小鼠肺纤维化，其机制可能与抑制肺组织炎症和TWEAK/Fn14信号通路有关。
Abstract:

Objective To observe the effects of Danshen Tongluo Jiedu decoction（DSTLD） on TWEAK/Fn14 pathway in mice with pulmonary fibrosis,and to investigate its effects and mechanism on pulmonary fibrosis in mice.Methods A total of fifty C57B/L mice were randomly divided into blank group,model group,positive control group1,25(OH)2D3 group, DSTLD group and DSTLD+TWEAK/ PDL192 group. The body mass of mice in each group was measured at 12 h after the last dose,then the BALF was collected,lung tissue was taken from the open chest,lung mass was measured,and the lung coefficients were calculated.The levels of IL-1β,TNF-α and IL-4 in BALF in the five groups were measured by enzyme marker.The histopathological changes of the lungs in the five groups were observed by HE staining and Masson′s trichrome staining.The mRNA expression levels of TWEAK,Fn14 and TGF-β1 were detected by RT-PCR.The levels of TNF-α,IL-1β,ICAM-1,VCAM-1,TWEAK,Fn14,TGF-β1,α-SMA,Smad3,p-Smad3,STAT3,p-STAT3 in lung tissue in the five groups were detected by Western blot.Results The lung coefficient in the model group, 1,25(OH)2D3 group and DSTLD group was significantly higher than that in the blank group (P<0.05);the lung coefficient in the DSTLD group and the DSTLD + PDL192 group was significantly lower than that in the model group (P<0.05);the lung coefficient in the DSTLD + PDL192 group was significantly lower than that in the 1,25(OH)2D3 group (P<0.05).The level of IL-1β,TNF-α and IL-4 in the BALF in the model group,1,25(OH)2D3 group and DSTLD group were significantly higher than those in the blank group (P<0.05),and the level of IL-1β in the BALF in the DSTLD + PDL192 group was significantly higher than that in the blank group (P<0.05).The levels of IL-1β,TNF-α and IL-4 in BALF in the 1,25(OH)2D3 group, DSTLD group and DSTLD + PDL192 group were significantly lower than those in the model group (P<0.05).The levels of IL-1β and IL-4 in the BALF in the DSTLD group and DSTLD + PDL192 group were significantly lower than those in the 1,25(OH)2D3 group (P<0.05).The levels of IL-1β and IL-4 in BALF in the DSTLD + PDL192 group were significantly lower than those in the DSTLD group (P<0.05).The levels of TWEAK,Fn14 and TGF-β1 mRNA in lung tissues in the model group and the level of TWEAK mRNA in lung tissues in the 1,25(OH)2D3 group were significantly higher than those in the blank group (P<0.05).The level of Fn14 mRNA in the lung tissue of mice in the DSTLD group and the of TWEAK,Fn14 and TGF-β1 mRNA in the lung tissue in the DSTLD + PDL192 group were significantly lower than those in the model group (P<0.05).The levels of TNF-α,IL-1β,ICAM-1 and VCAM-1 protein in the lung tissue in the model group and the levels of TNF-α and IL-1β protein in the lung tissue in the 1,25(OH)2D3 group and DSTLD group were significantly higher than those in the blank group (P<0.05);the levels of TNF-α and IL-1β in lung tissues in the DSTLD and DSTLD +PDL192 groups were significantly lower than those in the model group (P<0.05).The level of IL-1β protein in lung tissue in the DSTLD group and DSTLD + PDL192 group was significantly lower than that in the 1,25(OH)2D3 group (P<0.05).The level of IL-1β protein in lung tissue in the DSTLD + PDL192 group was significantly lower than that in the DSTLD group (P<0.05).The levels of TWEAK,Fn14,TGF-β1,α-SMA,p-Smad3,STAT3 and p-STAT3 protein in lung tissue in the model group were significantly higher than those in the blank group (P<0.05),and the levels of TWEAK,TGF-β1,α-SMA,p-Smad3,p-STAT3 and p-STAT3 protein in lung tissue in the 1,25(OH)2D3 group were significantly higher than those in the blank group (P<0.05),and the levels of TWEAK,α-SMA,p-Smad3,STAT3 protein were significantly higher than those in the blank group (P<0.05).The level of p-Smad3 protein in the lung tissue in the 1,25(OH)2D3 group was significantly lower than that in the model group (P<0.05),and the levels of TWEAK,Fn14,TGF-β1,α-SMA,p-Smad3 and STAT3 protein in the lung tissue in the DSTLD group and DSTLD + PDL192 group were significantly lower than those in the model group (P<0.05).The levels of TWEAK and STAT3 protein in lung tissue of mice in the DSTLD + PDL192 group were significantly lower than those in the 1,25(OH)2D3 group (P<0.05).Conclusion DSTLD can reduce bleomycin-induced pulmonary fibrosis in mice,and the mechanism may be related to the inhibition of lung tissue inflammation and TWEAK/Fn14 signaling pathway.


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更新日期/Last Update: 2021-12-05